TSA对牛成纤维细胞体外培养的影响  

Effect of TSA on bovine fibroblast cell in vitro

作　　者：岳顺利[1] 周佳勃[1,2] 周英[3] 于思淼 李有为[1] 孙铭菊 张利[1] 夏平[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [2]黑龙江省普通高等学校动物遗传育种与繁殖重点实验室,哈尔滨150030 [3]佳木斯大学医学院,黑龙江佳木斯154007

出　　处：《东北农业大学学报》2015年第6期67-72,共6页Journal of Northeast Agricultural University

基　　金：黑龙江省自然科学基金(留学回国)项目(LC2012C01);哈尔滨市科技创新人才项目(2013RFLXJ005);黑龙江省普通高等学校动物遗传育种与繁殖重点实验室开放课题(GXZDSYS-2012-05)

摘　　要：以培养到第三代的牛成纤维细胞为研究对象,经TSA处理后,进行细胞生长曲线绘制及形态、活力、周期的检测,并通过Real-Time PCR检测Oct4和Cdx2的表达情况。当TSA浓度高于50 nmol·L-1时,细胞出现异常形态,死细胞数目增多。MTT法证明浓度为50 nmol·L-1的TSA处理牛成纤维细胞24 h显著抑制细胞活力,但对细胞染色体整倍性形象差异不显著。与对照组相比,当TSA处理牛成纤维细胞48 h时,细胞染色体整倍率显著下降。50 nmol·L-1的TSA处理牛成纤维细胞24 h,60%以上细胞细胞周期阻滞在G0/G1期。Real-Time PCR结果表明与对照组相比TSA处理后,Oct4的表达量显著升高,而Cdx2基因的表达量则显著降低。结论,TSA处理后的牛成纤维细胞作为供体细胞,可能利于克隆胚胎表观重编程及发育。The third generation of cultured bovine fibroblasts for the study, such as cell growth curve, morphology, vitality, cell cycle, and the expression of Oct4 and Cdx2. The bovine fibroblast began to appear abnormal morphology and increase the number of dead cells when the concentration of TSA was higher than 50 nmol·L^-1. The result of MTT showed that the viability of bovine flbroblast was inhibited significantly after treatment by 50 nmol·L^-1 TSA for 24 h. Bovine fibroblast chromosomal composition was affected by HDACi. The diploid composition of the cells decreased significantly treated by 50 nmol·L^-1TSA for 48 h. The cell cycle was detected by flow cytometry showed that up to 60% of cells were arrested in G0/G1 phase treated by 50 nmol·L^-1 TSA for 24 h. Real-Time PCR results showed that compared with the control group, after TSA treatment, the expression of Oct4 increased significantly, while the expression level of Cdx2 gene decreased. HDACi treated bovine fibroblast cells may benefit the epigenetic reprogramming and the cloned embryos development when it was used as donor cells in somatic cell nuclear transfer.

关 键 词：组蛋白去乙酰化酶抑制剂 曲古霉素 牛成纤维细胞 体外培养 

分 类 号：S852.1[农业科学—基础兽医学]